Objective To investigate the cognition degree and clinical use of new COPD classification system of 2011 GOLD in respiratory specialists, and further analyze the reasons of failing to clinical use. Methods Respiratory specialists from 42 hospitals in Chongqing were investigated through questionnaire survey. The questionnaire contains two parts. The first part contains nine questions about the knowledge of 2011 GOLD new COPD classification system and its clinical use. The second part contains six questions about the reasons of failing to clinical use of the COPD classification system. Results A total of 204 valid questionnaires were recovered. More than 90% respiratory specialists had understood the new COPD classification system with different degree, and believed it is suitable for clinical use. More than twothirds respiratory specialists knew well the ways about CAT and mMRC, but only 24% specialists were using these ways. The main reasons of failing to clinical use were as follows: 60% specialists believed the pulmonary function test can evaluate the COPD classification, and 66. 7% specialists were limited by short visit time. The cognition degree and clinical use of the new COPD classification systemin the specialists from third grade A class hospitals was better than those from the other hospitals. But the difference was not significant among specialists with different professional title.Conclusion Respiratory specialists in Chongqing knew well about the new COPD classification systemin 2011 GOLD, but did not use it widely in clinical works due to the complicated operation of the new COPD classification system.
Objective To observe and analyze the clinical characteristics of children with autosomal dominant hereditary microcephaly with or without chorioretinopathy, lymphedema, or intellectual disability syndrome (MCLMR). MethodsA retrospective clinical study. In September 2023, the first patient and three family members (parents, brother) of MCLMR who were diagnosed through ophthalmic examination and genetic testing at Department of Ophthalmology of Henan Children's Hospital were included in the study. Clinical data were collected, inquired about medical history and family history in detail, and performed best corrected visual acuity (BCVA), optical coherence tomography (OCT), fluorescein angiography (FFA), flash visual evoked potential (F-VEP), full field electroretinogram (ERG), cranial magnetic resonance imaging (MRI), and systemic examination. 3 ml of peripheral venous blood were collected from the proband, her parents and younger brother, and extracted whole genome DNA. Second generation sequencing technology was used for gene sequencing. For suspected pathogenic sites, Sanger sequencing was used for validation, and bioinformatics analysis was performed to determine the pathogenicity of the genetic variant sites. The relevant literature of PubMed of the National Library of Medicine and Wan Fang Med Online by computer were searched. The genetic characteristics and conducted literature review were summarized. ResultsThe proband (Ⅱ-1) was an 8-year-old and 5-month-old female. Her head was relatively small, the lower jaw was small, the ears protrude, the nose was wide, the eyelid was tilted upwards, philtrum was long. Mild intellectual disability, no history of lymphedema. The BCVA values for the right and left eyes were 0.08 and 0.1, respectively. Bilateral nystagmus. Atrophic lesioned in the macular area and below choroid retina of both eyes. FFA examination showed mottled fluorescent staining in the macular area and the below retina, with no obvious fluorescein leakage in the late stage. OCT examination revealed shallow macular fovea morphology, absence of ellipsoidal bands, unclear layers, thinning of the entire retina, and significant atrophy of the choroid and retina beneath the macula. F-VEP examination, no waveform was detected in both eyes. Full field ERG examination showed severe reduction in amplitude of a wave and b wave in both eyes. Head magnetic resonance imaging showed widening of the subarachnoid space in the left temporal region, with no significant abnormal signals observed in the brain parenchyma. Her father (Ⅰ-1) had mild nystagmus and strabismus. The phenotypes of the eyes of the mother (Ⅰ-2) and brother (Ⅱ-2) were not significantly abnormal. The genetic testing results showed that the proband (Ⅱ-1) had a heterozygous missense mutation c.895A>G (p.Ile299Val) in exon 8 of the KIF11 gene, which was a known mutation. Her parents (Ⅰ-1, Ⅰ-2) and younger brother (Ⅱ-2) were both wild-type. The bioinformatics analysis results indicated that this mutation is a potentially pathogenic variant. A total of 109 cases were retrieved from 20 relevant literatures. Among them, 55 were male, 54 were females. There were 61 cases with family history and 48 cases without family history, respectively. Among the 109 cases, 98 cases (89.9%, 98/109) had microcephaly, 2 cases had premature closure of cranial sutures, and 11 patients underwent cranial MRI, which showed 11 cases of small head with simplified development of the cerebral gyrus. 50 cases (45.9%, 50/109) of lymphedema. 83 cases (76.1%, 83/109) of intellectual developmental disorders. 92 cases (84.4%, 92/109) had ocular abnormalities, 69 cases (63.3%, 69/109) had chorioretinopathy, 20 cases (18.3%, 20/109) had retinal folds, 10 cases (9.2%, 10/109) had nystagmus, and 17 cases (15.6%, 17/109) had retinal detachment. ConclusionsThe main clinical manifestations of MCLMR are microcephaly, chorioretinopathy, with or without lymphedema, and intellectual disability. The main manifestations of eye diseases are low vision, nystagmus, and chorioretinopathy. The heterozygous missense mutation c.895A>G (p.Ile299Val) in exon 8 of KIF11 gene is the pathogenic variant of this family.
Objective To investigate the regulatory role of PLA2G4A targeting in ferroptosis and its sensitizing effect on the ferroptosis inducer Erastin. Methods PLA2G4A expression in lung adenocarcinoma (LUAD) was assessed by analyzing data from The Cancer Genome Atlas and Clinical Proteomic Tumor Analysis Consortium databases, followed by immunohistochemical validation. PLA2G4A expression was knocked down in H1299 lung cancer cells using small interfering RNA. The correlation between PLA2G4A and ferroptosis marker genes was examined through gene correlation analysis and Western blotting. The regulatory relationship between PLA2G4A and ferrous ion (Fe2+) was analyzed using high-content fluorescence imaging. Cell proliferation after PLA2G4A inhibition and Erastin treatment was measured by CCK-8 assay. Flow cytometry and high-content fluorescence imaging were employed to evaluate the effects of PLA2G4A suppression combined with Erastin on intracellular Fe2+ and lipid peroxidation levels. Results Both mRNA (P<0.05) and protein (P<0.001) levels of PLA2G4A were significantly upregulated in LUAD tissues, and its high expression was associated with poor prognosis in LUAD patients (P<0.05). PLA2G4A expression was positively correlated with SLC7A11 expression (r=0.23, P<0.001). PLA2G4A knockdown suppressed SLC7A11 protein expression and increased cellular Fe2+ levels (P<0.01). Compared with the control group, PLA2G4A-silenced cells exhibited significantly reduced viability upon Erastin treatment (P<0.001). Furthermore, Erastin enhanced PLA2G4A targeting-induced Fe2+ accumulation and lipid peroxidation (P<0.001). Conclusion Targeting PLA2G4A induces ferroptosis in lung cancer cells by inhibiting SLC7A11 expression and enhances their sensitivity to Erastin.
ObjectiveTo summarize the research advances of pyroptosis in hepatic ischamia-reperfusion injury (IRI).MethodThe literatures about the studies of mechanism of pyroptosis in hepatic IRI were retrieved and analyzed.ResultsPyroptosis, also known as inflammatory necrocytosis, was proven to play an important role in the hepatic IRI. When hepatic ischemia-reperfusion occurred, the classical pathway of pyroptosis dependenting on caspase-1 and the non-classical pathway of pyroptosis dependenting on caspase-11 were initiated by specific stimulants, and leaded to the activation of gasdermin D, releases of proinflammatory factors such as interleukin-1β, interleukin-18, etc., and the recruitment and activation of neutrophils. Consequently, pyroptosis caused more severe hepatic inflammation and aggravated existing cell injury and dysfunction of liver during hepatic IRI.ConclusionsPyroptosis plays an important role in liver IRI. Further researches about mechanism of pyroptosis will be beneficial to the prevention and treatment of the pyroptosis of related diseases.
ObjectiveTo observe and analyze the clinical phenotype and genetic characteristics of COL2A1 and COL11A1 de novo mutation (DNM) related Stickler syndrome type Ⅰ and Ⅱ patients. MethodsA family-based cohort study. From December 2023 to November 2024, 4 patients (all probands) with Stickler syndrome diagnosed by clinical and genetic testing in Department of Ophthalmology of People's Hospital of Ningxia Hui Autonomous Region and their parents (8 cases) were included in the study. The patients came from 4 unrelated families. A detailed medical history was taken, and the patients underwent best-corrected visual acuity (BCVA), refraction, and fundus color photography examinations. Systemic examinations included the oral and facial regions, skeletal, joints, and hearing. Peripheral venous blood samples were collected from the patients and their parents, and genomic DNA was extracted. Whole-exome sequencing was used to screen for pathogenic genes and their loci, which were then validated by Sanger sequencing and combined with segregation analysis in the families to identify candidate gene mutation sites. The candidate variants were assessed for pathogenicity according to the American College of Medical Genetics and Genomics (ACMG) criteria and guidelines for the classification of genetic variants. Additionally, cross-species conservation analysis was performed to determine the evolutionary conservation of wild-type amino acids, and protein three-dimensional modeling techniques were used to characterize the spatial conformational changes of the variant proteins and the alterations in their local hydrogen bond networks. ResultsAmong the 4 patients, there were 2 males and 2 females; their ages ranged from 3 to 12 years. There were 2 cases of Stickler syndrome type Ⅰ (proband of families 1 and 2) and 2 cases of type Ⅱ (proband of families 3 and 4). The diopters ranged from ?8.00 to?18.00 D. BCVA ranged from no light perception to 0.6-. There were 2 cases each of vitreous membrane-like and “bead-like” opacity. Three cases showed peripapillary atrophy arcs and leopard pattern changes in the retina; one case had bilateral retinal detachment with a large macular hole in the left eye, which had previously been treated with vitrectomy surgery. One case had bilateral sensorineural hearing loss. There were 3 cases of simple micrognathia; one case had a flat nasal bridge, short nose, midface depression, and micrognathia. Two cases had excessive elbow joint extension. The phenotypes of the parents of the 4 patients were normal. Genetic testing results revealed that the probands of families 1 and 2 carried COL2A1 gene c.85+1G>C (M1) splice site variant and c.3950_3951insA (p.M1317Ifs*48) (M2) frameshift variant, respectively; the probands of families 3 and 4 carried COL11A1 gene (NM_001854.4) c.2549 G>T (p.G850V) (M3) missense variant and c.3816+6T>C (M4) splice site variant, respectively. The parents did not carry the related gene variants. Among them, M2, M3, and M4 are newly reported DNM. According to the ACMG guidelines, they were all considered likely pathogenic. The cross-species conservation analysis results showed that the wild-type amino acid of the COL11A1 gene M3 missense variant was highly conserved across multiple different species. Protein local structure modeling analysis revealed that the COL2A1 gene M2 frameshift variant and the COL11A1 gene M3 missense variant significantly altered the tertiary structure conformation of the protein, leading to abnormal spatial arrangement and hydrogen bond network in the key functional domains ConclusionThe COL2A1 gene M1 splice site variant, M2 frameshift variant, and the COL11A1 gene M3 missense variant, M4 splice site variant are respectively the potential pathogenic genes for families 1, 2, and families 3, 4; leading to the onset of Stickler syndrome type Ⅰ in families 1 and 2, and type Ⅱ in families 3 and 4.
Objective?To formulate an evidence-based conclusion concerning ultrasound screening for fetal malformations for a pregnant woman after 12 gestational weeks. Methods?Based on the clinical problem of whether pregnant women need ultrasound screening for fetal malformations after 11-14 gestational weeks, we used “ultrasound or sonography and prenatal or fetal at first trimester or 11-14 weeks; ultrasound exposure; fetal development” as the keywords and searched The Cochrane Library (Issue 4, 2008), MEDLINE (1981 to 2008), ACP Journal Club (1991 to 2008), and BMJ Clinical Evidence (1999 to 2008) for systematic reviews, randomized controlled trials (RCTs) and controlled clinical trials. The methodological quality of the included studies was assessed to identify the current best evidence. Results?Three systematic reviews, two RCTs and ten cohort studies were retrieved. The results showed ultrasound screening detected different fetal malformations in the first, second and third trimester. Not all of the fetal malformations could be detected through prenatal ultrasound screening. Nuchal translucency (NT) measurement as a tool for screening chromosomally abnormal fetuses and detecting fetal malformations by ultrasound proved to be effective if performed within 11-14 gestational weeks. The routine second trimester screening, however, could not be replaced by a detailed ultrasound examination at 11-14 gestational weeks. Most of the trials concluded that the effect of ultrasound on a fetus was not harmful. Conclusion?The findings of this study should reassure physicians and parents alike that ultrasound screening is an appropriate option for the pregnant women after 12 gestational weeks.
Methods To explore the anti-tumor effect of the zeste homolog enhancer 2 (EZH2) inhibitor EPZ011989 on retinoblastoma (RB) and its potential mechanism, and to evaluate whether it exerts an indirect neuroprotective effect on the optic nerve by regulating the epithelial-mesenchymal transition (EMT)-like process and the Wnt/β-catenin signaling pathway. MethodsWestern blotting (WB) was used to detect the expression of EZH2 protein in human retinal pigment epithelial cells (ARPE-19) and RB cells (Y-79, WERI-Rb-1). The half-maximal inhibitory concentration (IC50) of EPZ011989 on Y-79 and WERI-Rb-1 cells was determined using the Cell Counting Kit-8 (CCK-8). Y-79 and WERI-Rb-1 cells were treated with EPZ011989 at their respective IC50 concentrations and divided into untreated control groups (NC), dimethyl sulfoxide (DMSO) groups, and EPZ011989 treatment groups. Meanwhile, ARPE-19 cells were assigned to DMSO and EPZ011989 treatment groups. To verify the effect of EPZ011989 on the Wnt/β-catenin signaling pathway and its mechanism specificity, Y-79 and WERI-Rb-1 cells were respectively assigned to DMSO, EPZ011989 treatment, and EPZ011989 plus LiCl co-treatment groups. Cell proliferation, cell cycle, and apoptosis in different treatment groups were assessed by CCK-8 and flow cytometry, while WB was used to detect the expression of key proteins in the Wnt/β-catenin signaling pathway and EMT-related proteins.Animal experiment: 15 male C57BL/6J mice were randomly divided into 3 groups (n=5). A nude mouse RB subcutaneous transplantation tumor model was established by subcutaneous inoculation of Y-79 cells. The mice were orally administered with equal volume of DMSO (Control group), 100 mg/kg EPZ011989 (EPZ011989 low-dose group), and 300 mg/kg EPZ011989 (EPZ011989 high-dose group), twice a day for 4 weeks. Tumor volume and weight were monitored, and the expression levels of proliferation markers (Ki67) and EZH2 protein in the tumor-bearing tissues were detected by immunohistochemistry, and the expression levels of key proteins of the Wnt/β-catenin pathway in the tumor-bearing tissues were detected by WB. One-way analysis of variance was used for comparisons among multiple groups, and Tukey's post hoc test was used for pairwise comparisons between groups. ResultsThe expression levels of EZH2 protein in Y-79 and WERI-Rb-1 cells were significantly higher than those in ARPE-19 cells (P＜0.01). The IC50 values of EPZ011989 for Y-79 and WERI-Rb-1 cells were 1.851 μmol/L and 3.949 μmol/L, respectively. At these concentrations, EPZ011989 significantly inhibited the viability of both RB cell lines (P＜0.001) without significantly affecting the viability of ARPE-19 cells. Flow cytometry results showed that compared to the DMSO group, EPZ011989-treated Y-79 and WERI-Rb-1 cells exhibited consistent changes: the proportion of cells in the G0/G1 phase was significantly decreased (P＜0.001), while the proportions in the S and G2/M phases were significantly increased (P＜0.05); simultaneously, the apoptosis rates of both cell lines were significantly elevated (P＜0.001). Western blot analysis revealed that, compared to the DMSO group, EPZ011989 treatment significantly upregulated E-cadherin protein expression (P＜0.001) and downregulated N-cadherin and Vimentin expression (P＜0.001) in both RB cell lines, while also inhibiting the expression of key proteins in the Wnt/β-catenin pathway (Wnt1, β-catenin, c-MYC, Cyclin D1) (P＜0.01). In the rescue experiment with co-treatment of LiCl, compared with the EPZ011989 treatment group, the changes in cell cycle, increased apoptosis, inhibited Wnt/β-catenin pathway proteins (P＜0.001), and reversed EMT-like process-related proteins (down-regulation of E-cadherin, up-regulation of N-cadherin and Vimentin) caused by EPZ011989 were partially reversed in the EPZ011989+LiCl co-treatment group (P＜0.01). Tumor tissue detection showed that the tumor size in the high-dose EPZ011989 group was the smallest compared with the Control group and the low-dose EPZ011989 group. Compared with the Control group, the tumor volume and weight in the low-dose EPZ011989 group and the high-dose EPZ011989 group were significantly reduced (P < 0.001). Moreover, the reduction in tumor volume and weight in the high-dose EPZ011989 group was more significant (P＜0.001). The results of immunohistochemistry and WB assays showed that compared with the Control group, the expression of Ki67, EZH2, and key proteins of the Wnt/β-catenin pathway (Wnt1, β-catenin, c-MYC, Cyclin D1) in the low-dose EPZ011989 group and the high-dose EPZ011989 group were significantly decreased (P＜0.001), and the expression levels of these proteins in the high-dose group were the lowest. Conclusion: EPZ011989 inhibits the malignant progression of RB by suppressing the activation of the Wnt/β-catenin signaling pathway and may indirectly protect the function of the optic nerve by reducing the risk of optic nerve compression and regulating the EMT-like process and the Wnt/β-catenin signaling pathway. ConclusionEPZ011989 inhibits the malignant progression of RB by suppressing the activation of the Wnt/β-catenin signaling pathway, and may protect the optic nerve function indirectly by reducing the risk of optic nerve compression and regulating EMT-like processes and the Wnt/β-catenin signaling pathway.
Objective To investigate the correlation between OPRM1 A118G gene polymorphism and Eysenck personality type and pain sensitivity. Methods The surgical patients who were transferred from Department of Emergency Medicine to Department of General Surgery of Luzhou People’s Hospital between January 2018 and December 2020 were selected. Before surgery, Eysenck Personality Questionnai (EPQ) was used to investigate the patient’s personality type, and the pain threshold and pain tolerance threshold were determined by electric stimulation instrument. The OPRM1 A118G genotype of peripheral venous blood was detected by polymerase chain reaction-restriction fragment length polymorphism analysis technique. Patients were divided into wild homozygous (A/A) group, mutant heterozygous (A/G) group and mutant homozygous (G/G) group according to the typing results. The general condition, pain sensitivity, EPQ score, difference of Eysenck personality type and correlation between Eysenck personality type and pain sensitivity were analyzed. Results A total of 356 patients were enrolled, including 174 in A/A group, 136 in A/G group and 46 in G/G group. The mutation rate of OPRM1 A118G gene was 32.00%. There were statistically significant differences in pain sensitivity (pain threshold, pain tolerance threshold) and scores of introverted and extraverted, neurotic and dissemble personality types among three groups (P<0.05). There were significant differences in introverted and extraverted and psychotic personality types among the three groups (P<0.05). There were significant differences in pain threshold and pain tolerance threshold among different introverted, extraverted and psychotropic personality types (P<0.05). Conclusion Both OPRM1 A118G gene polymorphism and Eysenck personality type have influence on pain sensitivity, and there is a correlation between them.
Abstract： Objective To observe the significance of the changes of cell adhesion molecules (CAM) CD11b/CD18 and sPselectin during the perioperative period of open heart surgery under cardiopulmonary bypass (CPB), and investigate the roles of CD11b/CD18 and sPselectin in systemic inflammatory response triggered by CPB. Methods Thirty patients including 18 males and 12 females, age ranged from 29 to 55 years (45.3±8.1 years) having undergone valvular replacement for rheumatic heart disease in our hospital were selected as the subjects of this research. After anesthesia induction, radial arterial blood sample was collected at six different time points including the time prior to skin incision, and 30 min, 1 h, 6 h, 12 h and 24 h following the start of CPB. The expression levels of CD11b/CD18 were tested by flow cytometry, and concentration of sP-selectin in the plasma was measured with enzymelinked immunosorbent assay(ELISA). Results The expression of CD11b/CD18 was elevated at 30min after CPB, and it reached the peak (581.44±215.26) at 6 h after CPB with significant differences (Plt;0.05). Its expression started to drop at 12 h after CPB, but it was still higher than the expression level before CPB. The expression returned under the level before CPB at 24 h after CPB with insignificance differences (Pgt;0.05). The expression of sPselectin in the peripheral blood started to rise evidently at 30 min after CPB, reaching the peak (51.44±10.06 ng/ml) with significant differences (Plt;0.05). Its expression level decreased at 12 h after CPB and fell back below the level before CPB with insignificant differences (Pgt;0.05). Conclusion CPB can cause the expression of CD11b/CD18 and sPselectin to rise in the peripheral blood, which may play an important role in the systemic inflammatory response triggered by CPB.
Objective To investigate the controlled release effect and the anti-cancer cell ability of a 5-FU loaded poly-L-lactic acid?（PLLA） nanofibers membrane blending with keratin. Methods Making PLLA and keratin mix together and crosslinking to generate blending solution. Then the anti-cancer drug 5-FU was added into the solution to fabricate nanofibers membrane by high voltage electrospinning method. The micro morphology was observed by scanning electron microscope （SEM）. The controlled release effect of 5-FU from the nanofibers membrane was measured by high performance liquid chromatography （HPLC）. The cytotoxicity of 5-FU/PLLA keratin nanofibers membrane was evaluated by using 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide （MTT） assay on HCT116 cell lines. At the meantime, cell growth morphology of HCT116 in experiment group were observed by microscope and transmission electron microscope. Results 5-FU could be dispersed homogeneous in the PLLA/keratin nanofibers membrane through SEM. HPLC suggested that 5-FU could be diffused out from the fibers slowly and uniformly, which corresponded the zero order kinetics basically. After different treatment, the longer time the 5-FU/PLLA keratin nanofibers （experiment group） immerse in the medium, the much more swelling, apoptosis, and necrocytosis of the cells were observed. The cell viability for experiment group was （47.5±2.8）% by MTT, while the PLLA keratin nanofibers without 5-FU had no significant impact on cell viability （93.9±2.8）%, which was statistic significance （P＜0.01）. Conclusion 5-FU/PLLA keratin nanofibers membrane owns good controlled release effect and satisfies cell inhibitory effect against HCT116 cells in vitro，which suggested that it has a promising prospect for clinical therapy.
